Fungal genetic variability characterization in Clonostachys pytyrodes species using ITS-RFLP analysis: insights from bioinformatic análisis

PEDROZO, TT; BARENGO, MP; AMERIO, NS; ONETTO, AL; FASANO, MC; CORTESE, IJ; BICH, GA; ZAPATA, PD; CASTRILLO, ML

Simposio; 8VO SIMPOSIO ARGENTINO DE JOVENES INVESTIGADORES EN BIOINFORMÁTICA; 2023

Chemical pesticides, used for pest control, have negative consequences for the environment and human populations. As an alternative, the use of microorganisms or their metabolic products as biological control agents is a promising strategy. Thus, the correct identification of biotechnological potential fungal species in a bioproduct is a prerequisite for any biocontrol strategy. Clonostachys species are known for their potential as biocontrol. Understanding the genetic variability within Clonostachys populations is crucial for improving their accurate identification and harnessing their biotechnological potential. To achieve this, we first employed bioinformatics tools to retrieve 19 ITS sequences of selected Clonostachys species from the National Center for Biotechnology Information (NCBI) database. Next, we bioinformatically digested the retrieved sequences using the restriction enzymes AceIII, EcoRI, and BamHI to generate restriction fragment profiles and explore the genetic variability within the selected Clonostachys sequences. The bioinformatic analysis revealed a determined level of genetic diversity within the ITS region of Clonostachys species with enzymes EcoRI and BamHI. The enzyme AceIII did not cut any of the evaluated sequences. The virtual digestion of the ITS sequences using EcoRI and BamHI generated distinct restriction fragment profiles, indicating the presence of genetic variations among the Clonostachys isolates. BamHI consistently produced digestions that differentiated mainly C. pytyrodes from other closely related species. This finding suggests that ITS amplification combined with BamHI digestion could serve as a potential first molecular marker to detect C. pytyrodes from closely related species. The detection of species-specific digestion patterns through virtual enzyme digestion supports the utility of ITS-RFLP as a rapid technique for screening and monitoring Clonostachys populations. Overall, our study highlights the efficacy of combining bioinformatics and ITS-RFLP analysis to evaluate the genetic diversity within Clonostachys populations.